Urea Detection in Phosphate Buffer and Artificial Urine: A Simplified Kinetic Model of a pH-Sensitive EISCAP Urea Biosensor.

A simplified kinetic model for the quantitative analysis of a potentiometric, pH-based urea biosensor is presented. The device was an electrolyte-insulator-semiconductor capacitor (EISCAP) with a pH-sensitive Ta2O5 gate functionalized by a polyallylamine hydrochloride (PAH)/urease bilayer. Within the steady-state approximation, the kinetic equations yielded an implicit algebraic relation linking the bulk urea concentration to the local pH at the sensor surface. Numerical solution of this equation, combined with a fitting routine, provides the apparent Michaelis-Menten constant (KM) and the normalized maximum reaction rate (k¯V). Validation against the literature data confirmed the reliability of the approach. Experimental results were then analyzed in both phosphate buffer (PBS) and artificial urine (AU), covering urea concentrations of 0.1-50 mM. The fitted parameters showed comparable KM values of 10.9 mM (PBS) and 32.4 mM (AU), but strongly different k¯V values: 2.2×10-4 (PBS) versus 8.6×10-7 (AU). The three-order reduction in AU was attributed to the inhibitory effects inherent to complex biological fluids. These findings highlight the importance of the model-based quantitative analysis of EISCAP biosensors, enabling the accurate characterization of immobilized enzyme layers and guiding optimization for applications in realistic sample matrices.